CN101428041A - Cordyceps sinensis extract, preparation method and uses in preparing medicament - Google Patents

Cordyceps sinensis extract, preparation method and uses in preparing medicament Download PDF

Info

Publication number
CN101428041A
CN101428041A CNA2008100515981A CN200810051598A CN101428041A CN 101428041 A CN101428041 A CN 101428041A CN A2008100515981 A CNA2008100515981 A CN A2008100515981A CN 200810051598 A CN200810051598 A CN 200810051598A CN 101428041 A CN101428041 A CN 101428041A
Authority
CN
China
Prior art keywords
cordyceps
parts
extract
extraction
add
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CNA2008100515981A
Other languages
Chinese (zh)
Inventor
滕利荣
逯家辉
孟庆繁
刘丹梅
吴凌
付瑶
王迪
汤海峰
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jilin University
Original Assignee
Jilin University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Jilin University filed Critical Jilin University
Priority to CNA2008100515981A priority Critical patent/CN101428041A/en
Publication of CN101428041A publication Critical patent/CN101428041A/en
Pending legal-status Critical Current

Links

Landscapes

  • Medicines Containing Plant Substances (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention relates to a cordyceps extract and a preparation method thereof for a preparation, and belongs to the technical field of the extraction and the application of active components from a Chinese traditional medicine. The ultrasonic processing and the extraction method of combining alcohol extraction and water extraction are utilized. The extraction rate of the active components of polysaccharide can be increased by the ultrasonic processing, and since the alcohol extraction and the water extraction are combined, not only the extraction rate of the active components of the polysaccharide can be increased, but also the solvent is saved, the biological activity of the cordyceps polysaccharide is ensured, the utilization ratio of the raw material is increased to the utmost extent, the production cost is reduced, and the invention is suitable for the large-scale industrial production.

Description

A kind of Cordyceps extract and preparation method thereof and the application in the preparation medicine
Technical field:
The present invention relates to the preparation method of a kind of Cordyceps extract and preparation thereof, belong to Chinese medicine extracts active ingredients and applied technical field, also belong to field of traditional Chinese medicine pharmacy.
Background technology:
Cordyceps (Cordyceps militaris), another name Cordyceps militaris (L.) Link., Cordyceps militaris.Cordyceps militaris (L.) Link. belongs to Ascomycotina (Ascomycotina), Clavicipitaceae (Clavicipitaceae), Cordyceps (Cordyceps).Under the Cordyceps situation that supply falls short of demand, the Cordyceps militaris (L.) Link. equally celebrated for their achievements with Cordyceps is subjected to people's attention in the world day by day, and its exploitation and production have been developed new nutrient source and medicine source for body-care and field of medicaments." Xinhua's book on Chinese herbal medicine outline " record Cordyceps militaris (L.) Link. effects such as " property are flat, and sweet in the mouth has the lung benefiting kidney, mends marrow, hemostasis and phlegm " is used for diseases such as pulmonary tuberculosis, old man's weakness and anemia, is the Chinese crude drug of preciousness.Cordyceps militaris (L.) Link. has multiple pharmacological effect: can blood fat reducing, prevent arteriosclerosis, and the protection heart, cerebral tissue, tranquilizing soporific strengthens macrophage activity, anticancer, antiinflammatory, antibiotic, anti-hypoxia etc.; Cordyceps militaris (L.) Link. can be used for treating emphysema, prostate hyperplasia; Cordyceps militaris (L.) Link. can also human body immunity improving power, and auxiliary renal transplant recipients is got well, and increases sexual function etc.
The technology features of original capsule, tablet is to adopt the method for decocting in water precipitate with ethanol to extract multiple proteins and polysaccharide effective ingredient in the Cordyceps thalline Cordyceps thalline, and effective ingredient often extracts not exclusively.
Summary of the invention:
The object of the present invention is to provide a kind of Cordyceps extract, contain the effective active composition of Cordyceps.
The present invention also provides its preparation method, is applicable to suitability for industrialized production.
The present invention also provides the application of this Cordyceps extract in the preparation medicine, discloses corresponding drop pill, dispersible tablet and slow releasing tablet pharmaceutical dosage form.
Cordyceps extract of the present invention can obtain by the following method:
Get Cordyceps militaris (L.) Link. lyophilizing mycelium powder and 10-30% ethanol by 1:15-40 (W/V) mixed, 40-60 ℃ are carried out the ultrasound wave assisted extraction, and ultrasonic frequency is 20-40KHz, and the supersound process time is 30-90 minute, reprocessing 1-3 time, wherein concentration of alcohol is 10-30%; After extracting end, filter, merge repeatedly filtrate, get filtrate A;
Filter residue and drying with material-water ratio 1:30-1:50, in 40-60 ℃ of following supersound process (ultrasonic frequency is 20-40KHz) reaction 2-4 hours, gets liquor B;
Filtrate A is mixed with liquor B, and at 35-45 ℃, concentrating under reduced pressure under the 300-700mmHg adds 95% ethanol or dehydrated alcohol, makes that the ethanol weight content is 55-90% in the solution, leaves standstill 8-24 hour, utilizes freezer dryer to be dried to dry powder promptly.
Table 1 Different Extraction Method is to the influence of Cordyceps polysaccharide extraction ratio
Figure A200810051598D00051
Cordyceps drip pill method of the present invention may further comprise the steps:
With Cordyceps extract and substrate according to 1 part: the proportioning of (1~9) part, place heating while stirring in the heating container, standby until the fused solution that obtains containing drug extract and substrate, adjust the temperature control system of drop pill machine, make the temperature of drop pill machine reservoir remain on 50 ℃~90 ℃, the temperature of condensing agent remains on 5 ℃~20 ℃, when treating that the temperature of condensing agent in drop pill machine reservoir and the condensation column is stable respectively and reaching desired state of temperature, the fused solution that will contain Cordyceps extract and substrate places in the reservoir of drop pill machine, splashes into molding in the condensing agent.
Cordyceps dispersible tablet method of the present invention may further comprise the steps:
Cordyceps extract and adjuvant are pulverized, cross 80~200 mesh sieves, get 10~30 parts of Cordyceps extracts, add 5~70 parts disintegrating agent, 10~85 parts filler, the wet adhesive that adds 5~30 parts and make suitable soft material, cross the 20-40 mesh sieve and granulate, under 40~80 ℃ temperature, dry, 20-40 mesh sieve granulate adds 1~3 part lubricant, mix homogeneously in the dried granule that makes, tabletting promptly, the preferred tablet Hardness Control is at 4-7kg.
Cordyceps slow releasing tablet preparation method of the present invention may further comprise the steps:
Get 20.0~60.0 parts Cordyceps extract, add 10.0~70.0 parts slow-release material and 5.0~50.0 parts filler, mixing is crossed 80~120 mesh sieves after crushed; Add suitable amount of adhesive, adopt dry method or wet granulation, granulate, add 1.0~3.0 parts lubricant mixing, the tablet machine tabletting is made the slow releasing tablet that contains effective principal agent amount and is got final product.
It is as follows that the existing relatively Cordyceps extraction of effective components thing of the present invention is compared the good effect that has: used ultrasonic Treatment, alcohol extraction to carry the extracting method that combines with water, supersound process can improve the extraction of effective ingredient, alcohol extraction is carried combining with water and both can be improved extraction ratio of effective constituents, can save solvent again, thereby guaranteed the biological activity of Cordyceps polysaccharide, increase utilization ratio of raw materials to greatest extent, reduced production cost, be fit to large-scale industrial production.
The specific embodiment:
Embodiment 1
Get Cordyceps militaris (L.) Link. lyophilizing mycelium powder 100g, in ratio adding 10 ~ 30% ethanol mixing of 1: 40 (W/V), 40 ℃ are carried out the ultrasound wave assisted extraction, ultrasonic frequency is 40KHz, the supersound process time is 60 minutes, reprocessing 3 times, and wherein concentration of alcohol is respectively 10%, 20%, 30%; After extracting end, filter, merge repeatedly filtrate, get filtrate A;
Filter residue and drying, with material-water ratio 1:30, in 40 ℃ of following supersound process, ultrasonic frequency is 30KHz, reacts 4 hours, gets liquor B;
Filtrate A is mixed with liquor B, and at 35-45 ℃, concentrating under reduced pressure under the 700mmHg adds 95% ethanol or dehydrated alcohol, makes that the ethanol weight content is 55% in the solution, leaves standstill 24 hours, and water solublity Cordyceps militaris (L.) Link. crude polysaccharides is separated out; Utilize freezer dryer to be dried to dry powder promptly.
Embodiment 2
Get ratio adding 10 ~ 30% ethanol mixing of Cordyceps militaris (L.) Link. lyophilizing mycelium powder 100g in 1: 15 (W/V), 60 ℃ are carried out the ultrasound wave assisted extraction, and ultrasonic frequency is 20-40KHz, and the supersound process time is 90 minutes, reprocessing 3 times, wherein concentration of alcohol is for being respectively 10%, 20%, 30%; After extracting end, filter, merge repeatedly filtrate, get filtrate A;
Filter residue and drying, with material-water ratio 1:50, in 60 ℃ of following supersound process, ultrasonic frequency is 40KHz, reacts 3 hours, gets liquor B;
Filtrate A is mixed with liquor B, and at 35-45 ℃, concentrating under reduced pressure under the 300mmHg adds 95% ethanol or dehydrated alcohol, makes that the ethanol weight content is 90% in the solution, leaves standstill 12 hours, and water solublity Cordyceps militaris (L.) Link. crude polysaccharides is separated out; Utilize freezer dryer to be dried to dry powder promptly.
Embodiment 3: cordyceps drip pill
1. Cordyceps extract: get embodiment 1 and be prepared into Cordyceps extract.
2. preparation method: get 100 parts of Cordyceps extracts, with 100 parts of PEG4000 mixings, place heating while stirring in the heating container, standby until the fused solution that obtains containing drug extract and substrate, adjust the temperature control system of drop pill machine, make the reservoir heating of drop pill machine and remain on 70 ℃~80 ℃, the temperature of condensing agent remains on 5 ℃~8 ℃, when treating that the temperature of condensing agent in drop pill machine reservoir and the condensation column is stable respectively and reaching desired state of temperature, the fused solution that will contain Cordyceps extractum and substrate places in the reservoir of drop pill machine, splashes into molding in the condensing agent dimethicone.
Embodiment 4:
1. Cordyceps extract: get embodiment 2 and be prepared into Cordyceps extract.
2. preparation method: get 100 parts of Cordyceps extracts, with 300 parts of pools husky nurse 188 mixings that fall, place heating while stirring in the heating container, standby until the fused solution that obtains containing drug extract and substrate, adjust the temperature control system of drop pill machine, make the reservoir heating of drop pill machine and remain on 70 ℃~80 ℃, the temperature of condensing agent remains on 5 ℃~8 ℃, when treating that the temperature of condensing agent in drop pill machine reservoir and the condensation column is stable respectively and reaching desired state of temperature, the fused solution that will contain Cordyceps extractum and substrate places in the reservoir of drop pill machine, splashes into molding in the condensing agent dimethicone.
Embodiment 5: the Cordyceps dispersible tablet
Prescription: composition (weight percent content)
20 parts of Cordyceps extracts, 32 parts of starch, 15 parts of sodium carboxymethyl cellulose, 10 parts of crosslinked carboxymethyl feculas, 15 parts of microcrystalline Cellulose, 2 parts of acesulfame-Ks, 5 parts of 30 POVIDONE K 30 BP/USP 30 ethanol, 1 part of micropowder silica gel;
Get Cordyceps extract and the adjuvant in the above-mentioned prescription that embodiment 1 makes and cross 100 mesh sieve pulverization process, again with Cordyceps extract and sodium carboxymethyl cellulose, crosslinked carboxymethyl fecula sodium, microcrystalline Cellulose, the abundant mix homogeneously of starch, 30 POVIDONE K 30 BP/USP 30 alcoholic solution of employing 5% are made soft material as wet adhesive with above-mentioned solid mixture, 20 mesh sieves are granulated in 60 ℃ of oven dry, 20 mesh sieve granulate add the micropowder silica gel mixing, the control tablet weight, tabletting promptly gets the Cordyceps dispersible tablet.
The dispersible tablet uniformity inspection that embodiment obtains: in 20 ± 1 ℃ of water, all disintegrates and the time by No. 2 sieves are not more than 140 seconds, and is up to specification.
Inspection method about the dispersing uniformity of dispersible tablet of the present invention and dissolution is as follows.
1. the inspection of the dispersing uniformity of dispersible tablet: by the requirement inspection down of 2005 editions two appendix IA dispersible tablets of Chinese Pharmacopoeia item; get 2 of dispersible tablets of the present invention; place the jolting of 100ml water; in 20 ± 1 ℃ of water; all disintegrate was by No. 2 sieves in 3 minutes, and the employing device can be by the requirement inspection of apparatus under 2005 editions two appendix XA tablets of the Chinese Pharmacopoeia item.
The mensuration of dispersing uniformity of the present invention:
The dispersing uniformity check result of table 2. Cordyceps dispersible tablet
Figure A200810051598D00071
2. the dissolution test of dispersible tablet: get this product, carry out according to two appendix dissolution determination second methods of Chinese Pharmacopoeia version in 2005.Rotating speed is 100r/min, and medium is a 900ml degassing deionized water, and temperature is 37 ± 0.5 ℃, and timing sampling 5ml filters through microporous filter membrane (0.45um), replenishes the deionized water of equal volume and uniform temp simultaneously.Subsequent filtrate is measured polysaccharide concentration through the sulphuric acid anthrone method, calculates the cumulative release amount, and limit is 80% of a labelled amount, should be up to specification.
The dissolution of table 3. Cordyceps dispersible tablet and common Cordyceps tablet relatively
Figure A200810051598D00072
By table 1,2 test data as can be seen, the complete conformance with standard of disintegrate dispersive property of Cordyceps dispersible tablet, dissolving out capability obviously is better than ordinary tablet.
Embodiment 6 Cordyceps slow releasing tablet
Prescription: 150.0 parts of Cordyceps extracts, 80.0 parts of hydroxypropyl methylcellulose, 67.0 parts of ethyl celluloses,
50.0 parts of microcrystalline Cellulose, 3.0 parts of magnesium stearate, 20 parts of 50% ethanol
Preparation method: take by weighing the Cordyceps extract of described deal, add the slow-release material and the filler of described deal, mixing is all pulverized the back and is crossed 80~120 mesh sieves; Add suitable amount of adhesive, adopt wet granulation, with 18 mesh sieve granulate, 40~60 ℃ of dryings were taken out after 1~2 hour, added the lubricant mixing of described deal, and the tablet machine tabletting is made the slow releasing tablet that sheet weighs 0.5g, promptly.
Embodiment 7
Prescription: 150.0 parts of Cordyceps extracts, 60.0 parts of hydroxypropyl methylcellulose, 45.0 parts of methylcellulose,
92.0 parts of microcrystalline Cellulose, 3.0 parts of magnesium stearate, 20 parts of 50% ethanol
Preparation method: take by weighing the Cordyceps extract of described deal, add the slow-release material and the filler of described deal, mixing is all pulverized the back and is crossed 80~120 mesh sieves; Add suitable amount of adhesive, adopt wet granulation, with 18 mesh sieve granulate, 40~60 ℃ of dryings were taken out after 1~2 hour, added the lubricant mixing of described deal, and the tablet machine tabletting is made the slow releasing tablet that sheet weighs 0.5g, promptly.
Embodiment 8
Prescription: 150.0 parts of Cordyceps extracts, hyprolose, 40.0 parts, 25.0 parts of polyvidones,
132.0 parts of lactose, 3.0 parts of magnesium stearate, 20 parts of 50% ethanol
Preparation method: take by weighing the Cordyceps extract of described deal, add the slow-release material and the filler of described deal, mixing is all pulverized the back and is crossed 80~120 mesh sieves; Add suitable amount of adhesive, adopt wet granulation, with 18 mesh sieve granulate, 40~60 ℃ of dryings were taken out after 1~2 hour, added the lubricant mixing of described deal, and the tablet machine tabletting is made the slow releasing tablet that sheet weighs 0.5g, promptly.

Claims (4)

1, a kind of Cordyceps extract can obtain by the following method:
Get the alcoholic acid mixed proportion of Cordyceps militaris (L.) Link. lyophilizing mycelium powder and 10-30% and count 1:15 ~ 40 by (W/V), 40 ~ 60 ℃ are carried out the ultrasound wave assisted extraction, and ultrasonic frequency is 20 ~ 40KHz, and the supersound process time is 30 ~ 90 minutes, reprocessing 1-3 time, wherein concentration of alcohol is 10 ~ 30%; After extracting end, filter, merge repeatedly filtrate, get filtrate A;
Filter residue and drying is with material-water ratio 1:(30 ~ 50), in 40 ~ 60 ℃ of following supersound process, ultrasonic frequency is 20-40KHz, reacts 2-4 hours, gets liquor B;
Filtrate A is mixed with liquor B, and at 35-45 ℃, concentrating under reduced pressure under the 300-700mmHg adds 95% ethanol or dehydrated alcohol, makes that the ethanol weight content is 55-90% in the solution, leaves standstill 8-24 hour, utilizes freezer dryer to be dried to dry powder promptly.
2, the drop pill made of the described Cordyceps extract of claim 1 is characterized in that:
With claim 1 Cordyceps extract and substrate according to 1 part: the weight proportion of (1~9) part, place that to be heated to fusion in the heating container while stirring standby, the temperature of adjusting drop pill machine reservoir remains on 50 ℃~90 ℃, the temperature of condensing agent remains on 5 ℃~20 ℃, the fused solution that will contain Cordyceps extractum and substrate places in the reservoir of drop pill machine, splashes into molding in the condensing agent.
3, the dispersible tablet made of the described Cordyceps extract of claim 1 is characterized in that:
With claim 1 Cordyceps dry extract and adjuvant drying, pulverizing, cross 80~200 mesh sieves, get 10~30 parts of Cordyceps extracts, add 5~70 parts disintegrating agent, 10~85 parts filler, 5~10 parts of wet adhesives and make soft material, cross the 20-40 mesh sieve and granulate, under 40 ~ 80 ℃ temperature, dry, 20-40 mesh sieve granulate, the lubricant that adds 1~3 part in the dried granule that makes, mix homogeneously, tabletting are promptly.
4, the slow releasing tablet made of the described Cordyceps extract of claim 1 is characterized in that:
Get 20.0~60.0 parts Cordyceps extract drying and crushing, add 10.0~70.0 parts slow-release material and 5.0~50.0 parts filler, mixing is crossed 80~120 mesh sieves after crushed; Add suitable amount of adhesive, adopt dry method or wet granulation, granulate, add 1.0~3.0 parts lubricant mixing, the tablet machine tabletting gets final product.
CNA2008100515981A 2008-12-12 2008-12-12 Cordyceps sinensis extract, preparation method and uses in preparing medicament Pending CN101428041A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CNA2008100515981A CN101428041A (en) 2008-12-12 2008-12-12 Cordyceps sinensis extract, preparation method and uses in preparing medicament

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNA2008100515981A CN101428041A (en) 2008-12-12 2008-12-12 Cordyceps sinensis extract, preparation method and uses in preparing medicament

Publications (1)

Publication Number Publication Date
CN101428041A true CN101428041A (en) 2009-05-13

Family

ID=40643809

Family Applications (1)

Application Number Title Priority Date Filing Date
CNA2008100515981A Pending CN101428041A (en) 2008-12-12 2008-12-12 Cordyceps sinensis extract, preparation method and uses in preparing medicament

Country Status (1)

Country Link
CN (1) CN101428041A (en)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101766662B (en) * 2010-01-07 2012-05-23 南京中科药业有限公司 Process for efficiently concentrating cordycepin
CN102499435A (en) * 2011-11-02 2012-06-20 深圳市大百汇技术有限公司 Cordyceps militaris extraction method and cigarette containing cordyceps militaris extract
CN103432172A (en) * 2013-08-23 2013-12-11 周秋火 Cordyceps sinensis tablets
WO2017084029A1 (en) * 2015-11-17 2017-05-26 罗明镜 Method for extracting cordyceps sinensis extract
WO2017084030A1 (en) * 2015-11-17 2017-05-26 罗明镜 Method for extracting cordyceps sinensis extract
CN113546097A (en) * 2021-07-21 2021-10-26 广东省科学院微生物研究所(广东省微生物分析检测中心) Application of ethanol extract of cordyceps guangdongensis fruiting body in preparation of medicine for preventing obesity and hyperlipidemia

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101766662B (en) * 2010-01-07 2012-05-23 南京中科药业有限公司 Process for efficiently concentrating cordycepin
CN102499435A (en) * 2011-11-02 2012-06-20 深圳市大百汇技术有限公司 Cordyceps militaris extraction method and cigarette containing cordyceps militaris extract
CN102499435B (en) * 2011-11-02 2014-01-01 深圳市大百汇技术有限公司 Cigarette containing cordyceps militaris extract
CN103432172A (en) * 2013-08-23 2013-12-11 周秋火 Cordyceps sinensis tablets
WO2017084029A1 (en) * 2015-11-17 2017-05-26 罗明镜 Method for extracting cordyceps sinensis extract
WO2017084030A1 (en) * 2015-11-17 2017-05-26 罗明镜 Method for extracting cordyceps sinensis extract
CN113546097A (en) * 2021-07-21 2021-10-26 广东省科学院微生物研究所(广东省微生物分析检测中心) Application of ethanol extract of cordyceps guangdongensis fruiting body in preparation of medicine for preventing obesity and hyperlipidemia

Similar Documents

Publication Publication Date Title
CN101167749B (en) Preparing method for improving toad venom taste
CN101428041A (en) Cordyceps sinensis extract, preparation method and uses in preparing medicament
CN103655639A (en) Spirulina tablet, and preparation method thereof
JP2016539955A (en) Drug composition, method for producing the same, and use
CN102772440A (en) Processing method of ultra-micro wall-breaking oral-tablet decoction pieces by traditional Chinese medicines
CN102085344B (en) Aplotaxis carminative sustained-release preparation and preparation method thereof
CN101401796A (en) Pramipexole orally disintegrating tablets and preparation method thereof
CN104906160B (en) A kind of enteric coated preparations of erigeron breviscapus extract
CN102091219A (en) New Huang's sound pills and preparation method thereof
CN100528143C (en) Method of preparing concentrated pill of six ingredients with rehmannia
CN1329062C (en) Method for preparing medicine for treating cataclasis and soft tissue injury
CN103381193A (en) Rhodiola rosea micro powder tablet and preparation method thereof
CN101095709A (en) Panax notoginseng floral leaf effervescent tablets and method for preparing the same
CN101732555A (en) Medicament for treating cold cough and chronic bronchitis
CN102228484A (en) Chinese medicinal preparation Jinshuibao oral liquid and preparation method thereof
CN104069347A (en) Preparation method of polygonatum odoratum total saponin dispersible tablet
CN102772439A (en) Processing method of ultra-micro wall-breaking capsule-preparation decoction pieces by traditional Chinese medicines
CN100450504C (en) Medication for treating pharyngitis
CN101766683B (en) Salvia dispersible tablet and application thereof
CN105497722A (en) Preparation method for Mongolian four-species elecampane inula root decoction superfine powder tablets, granules and capsules
CN100444846C (en) Dispersion tablet contg. Gynostemma petaphylla total glucosides, and its prepn. method
CN100358554C (en) Chinese medicinal capsules for disinfecting and relieving toxicity and their preparation
CN101011383A (en) Dispersible tablet of coumarin derivative and its preparation method
CN100415269C (en) Method for preparing cow-bezoar snake bile Sichuan fritillary bulb dispersive tablet
CN102552383A (en) Compound jellyfish antihypertensive dispersible tablet and preparation method thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C12 Rejection of a patent application after its publication
RJ01 Rejection of invention patent application after publication

Open date: 20090513